We report the isolation of an alternatively spliced human lysosome-associated membrane protein-2 (h-lamp-2) transcript which is overexpressed in human muscle. The cloning of this transcript is an indication for the tissue-specific expression of lysosomal membrane proteins and implicates the possibility of multiple functions for the protein products of the h-lamp-2 gene, as well as other lysosome-associated membrane proteins. The new transcript, designated h-lamp-2b, results from the alternative splicing of the last exon, exon 9, the alternative form of which is approximately 2800 bp in length. The resulting protein is identical in length to the previously reported h-lamp-2 protein, 410 amino acids including the leader peptide. This final exon, which encodes the last eleven amino acids of the luminal domain, the 24 amino acid transmembrane spanning region, and an eleven amino acid cytoplasmic tail, shows complete conservation of the Gly.Tyr.X.X lysosomal targeting signal with regard to its position relative to the transmembrane spanning region and the carboxy terminus of the protein. Immune electron microscopy studies verified localization of this alternative gene product to the lysosomal membrane.